Molecular Identification of Aminoglycoside-Modifying Enzymes and Plasmid-Mediated Quinolone Resistance Genes among Klebsiella pneumoniae Clinical Isolates Recovered from Egyptian Patients

Table 2

Primer sets and PCR cycling conditions used for amplification of genes encoding quinolone resistance.

Prime	Sequence	Gene	Reference	Amplification conditions	Amplicon size (bp)
QnrA	F: AGAGGATTTCTCACGCCAGG; R: TGCCAGGCACAGATCTTGAC	qnrA	[23]	Initial denaturation at 95°C for 15 min, then 30 cycles of 95°C for 1 min, 55°C for 1 min, and 72°C for 5 minutes, and one cycle of final elongation at 72°C.	580
QnrB	F: GGMATHGAAATTCGCCACTG; R: TTTGCYGYYCGCCAGTCGAA	qnrB	[23]		264
QnrC	F: GGGTTGTACATTTATTGAATCG; R: CACCTACCCATTTATTTTCA	qnrC	[24]		307
QnrD	F: CGAGATCAATTTACGGGGAATA; R: AACAAGCTGAAGCGCCTG	qnrD	[24]	Initial denaturation at 95°C for 15 min, then 30 cycles of 95°C for 1 min, 56°C for 1 min, and 72°C for 5 minutes, and one cycle of final elongation at 72°C.	533
QnrS	F: GCAAGTTCATTGAACAGGGT; R: TCTAAACCGTCGAGTTCGGCG	qnrS	[23]		428
ACC (6′)-Ib-cr	F: TTGCGATGCTCTATGAGTGGCTA; R: CTCGAATGC-CTGGCGTGTTT	acc (6′)-Ib-cr	[25]		482
OqxA	F: CTCGGCGCGATGATGCT; R: CCACTCTTCACGGGAGACGA	oqxA	[26]	Initial denaturation at 95°C for 15 min, then 30 cycles of 95°C for 1 min, 60°C for 1 min, and 72°C for 5 minutes, and one cycle of final elongation at 72°C.	392
OqxB	F: TTCTCCCCCGGCGGGAAGTAC; R: CTCGGCCATTTTGGCGCGTA	oqxB	[26]	Initial denaturation at 95°C for 15 min, then 35 cycles of 95°C for 1 min, 58°C for 1 min, and 72°C for 5 minutes, and one cycle of final elongation at 72°C.	512
Qep	F: AACTGCTTGAGCCCGTAGAT; R: GTCTACGCCATGGACCTCAC	qepA	[27]	Initial denaturation at 95°C for 15 min, then 30 cycles of 95°C for 1 min, 55°C for 1 min, and 72°C for 5 minutes, and one cycle of final elongation at 72°C.	596

qnr: quinolone resistance protein, acc(6′)-Ib-cr: aminoglycoside acetyl transferase-ciprofloxacin variant, Oqx: olaquindox, and Qep: quinolone efflux pump.